Optimization of affinity capillary electrophoresis (ACE) conditions using chemometrics

ANYL 139

Ruth E. Montes, beberuth@gmail.com1, Amaris Pao1, Frank A. Gomez, fgomez2@calstatela.edu2, and Grady S. Hanrahan, ghanrah@calstatela.edu3. (1) Chemistry & Biochemistry, California State University, Los Angeles, 5151 State University Dr, Los Angeles, CA 90032, (2) Chemistry and Biochemistry, California State University, Los Angeles, 5151 State University Drive, Los Angeles, CA 90032, (3) Department of Chemistry & Biochemistry, California State University, Los Angeles, 5151 State University Drive, Los Angeles, CA 90032
Affinity capillary electrophoresis (ACE) is a versatile technique to estimate binding parameters between receptors and ligands. One variation in ACE we have pioneered is flow-through partial filling (FTPFACE). In this method, the capillary is first partially filled with a zone of ligand followed by a plug containing receptor and markers. Upon application of voltage the plug flows into the ligand zone where a dynamic equilibrium is established between receptor and ligand. The analysis of the receptor's electrophoretic mobility (µ), relative to markers, as a function of the ligand concentration yields a value for the binding constant. Chemometrics allows for factor screening and optimization of experimental conditions. For the first time its use, in particular fractional factorial and Box-Behnken designs, is integrated in ACE analysis. Using the model system carbonic anhydrase B (CAB, E.C.4.2.1.1) and its binding to an arylsulfonamide, parameters including injection time, voltage, and capillary length are examined.
 

General Papers
7:00 PM-9:00 PM, Sunday, 10 September 2006 Moscone Center -- Hall D, Poster

Division of Analytical Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006